Recombinant Human CFAP77 Protein: Research Guide

Recombinant Human CFAP77 Protein is a cilia- and flagella-associated recombinant protein intended for in vitro research applications including functional studies, antibody validation, and structural investigations of axonemal components.

Product Name

Recombinant Human CFAP77 Protein

SKU

rp374838

Protein Family

Cilia and Flagella Associated Protein (CFAP) family

Species Origin

Human (Homo sapiens)

Protein Type

Recombinant protein expressed in a heterologous system

Primary Application

In vitro research: functional assays, antibody validation, binding studies

Storage Recommendation

Store at -80°C; avoid repeated freeze-thaw cycles (general guideline for recombinant proteins)

Biological Background of CFAP77

CFAP77 (Cilia and Flagella Associated Protein 77) is a member of the broader CFAP protein family, a group of proteins that localize to cilia and flagella and contribute to the structural integrity and motility of these organelles. Cilia and flagella are microtubule-based appendages found on the surface of many eukaryotic cells, playing critical roles in fluid movement, sensory signaling, and cell motility.

The axoneme, the core structural unit of cilia and flagella, is composed of highly organized microtubule doublets surrounded by an array of associated proteins. CFAP77 is thought to associate with axonemal structures, although its precise molecular function requires further characterization. Disruption of cilia-associated proteins has been linked to a class of diseases termed ciliopathies, which can affect multiple organ systems including the respiratory tract, reproductive system, and central nervous system.

Recombinant production of CFAP77 enables researchers to study this protein in a controlled, cell-free environment, facilitating biochemical and biophysical investigations that would otherwise be challenging using endogenous protein sources.

Recommended Research Applications

Recombinant Human CFAP77 Protein (rp374838) is suitable for a range of in vitro research applications. Researchers should validate the protein for each specific application prior to use in critical experiments.

• Antibody Validation: Use as a positive control antigen for Western blot, ELISA, or immunoprecipitation when validating anti-CFAP77 antibodies.
• Protein–Protein Interaction Studies: Employ in pull-down assays or co-immunoprecipitation experiments to identify binding partners within axonemal or ciliary protein networks.
• Structural and Biophysical Studies: Suitable as a substrate for circular dichroism, dynamic light scattering, or other biophysical characterization techniques.
• Functional Assays: Investigate the biochemical activity or binding properties of CFAP77 under defined in vitro conditions.
• Standard Curve Generation: Use as a quantitative standard in ELISA-based detection systems targeting CFAP77.

Note: All applications should be empirically validated by the end user. Specific activity and suitability for particular assay formats depend on lot-specific characterization data available from Aladdin Scientific upon request.

Handling, Reconstitution, and Storage Guidelines

Proper handling of Recombinant Human CFAP77 Protein is essential to preserve its integrity and biological activity. The following are general best-practice guidelines applicable to recombinant proteins; always refer to the product-specific datasheet for lot-confirmed parameters.

Upon Receipt

• Inspect the shipment immediately upon arrival. Check that the product has been maintained at the correct temperature during transit.
• Centrifuge the vial briefly (1,000–2,000 × g, 30 seconds) before opening to consolidate any material on the cap or walls.

Reconstitution

• If supplied lyophilized, reconstitute in the buffer recommended on the datasheet (typically sterile PBS or manufacturer-specified buffer) to the indicated concentration.
• Allow the vial to equilibrate to room temperature before reconstitution to minimize aggregation risk.
• Mix gently by pipetting; avoid vortexing, which may cause protein denaturation.

Storage

• Store at -80°C for long-term stability.
• For short-term use (up to 1 week), storage at 4°C may be acceptable; confirm with datasheet specifications.
• Prepare single-use aliquots prior to freezing to avoid repeated freeze-thaw cycles, which can degrade protein activity and increase aggregation.
• Addition of a carrier protein (e.g., 0.1% BSA) may improve stability during extended storage at low concentrations.

Quality Considerations and Experimental Best Practices

To obtain reproducible and reliable results with Recombinant Human CFAP77 Protein, consider the following quality and experimental best-practice points:

• Purity Assessment: Confirm protein purity by SDS-PAGE or equivalent method prior to critical experiments. Contact Aladdin Scientific for lot-specific Certificate of Analysis (CoA) data.
• Endotoxin Levels: For cell-based assays, verify that endotoxin levels are within acceptable limits. Request endotoxin testing data from the product datasheet or CoA.
• Protein Concentration Determination: Measure protein concentration by a validated method (e.g., BCA assay, A280 absorbance) rather than relying solely on nominal concentration, as slight variations can occur between lots.
• Positive and Negative Controls: Always include appropriate controls in functional and binding assays to account for non-specific signals.
• Aggregation Monitoring: If protein turbidity is observed after reconstitution or thawing, centrifuge at ≥10,000 × g for 10 minutes at 4°C and use the clarified supernatant.
• Lot Consistency: When reproducibility across experiments is critical, reserve sufficient aliquots from a single lot to complete a project.

Frequently asked questions

What is CFAP77 and why is it relevant to cilia research?

CFAP77 (Cilia and Flagella Associated Protein 77) is a protein that associates with the ciliary and flagellar axoneme. It is of interest in cilia biology research because disruption of axonemal proteins is linked to ciliopathies, a group of disorders affecting organs dependent on cilia function such as the lungs, kidneys, and reproductive system. Recombinant CFAP77 enables in vitro investigation of its function and interactions.

How should I reconstitute and store Recombinant Human CFAP77 Protein to maintain activity?

If the protein is supplied in lyophilized form, reconstitute it in the buffer specified on the product datasheet, gently mixing by pipetting without vortexing. For long-term storage, keep aliquots at -80°C and avoid repeated freeze-thaw cycles. For short-term use, storage at 4°C for up to one week may be acceptable; always consult the lot-specific datasheet for confirmed parameters.

Which experimental applications is Recombinant Human CFAP77 Protein validated for?

Recombinant Human CFAP77 Protein is intended for in vitro research applications including antibody validation (Western blot, ELISA), protein–protein interaction studies, biophysical characterization, and use as an ELISA standard. Users should empirically validate the protein for each intended application, and lot-specific characterization data can be requested from Aladdin Scientific.

How do I minimize protein aggregation when working with CFAP77?

To minimize aggregation, allow the vial to reach room temperature before reconstitution, mix gently without vortexing, and prepare single-use aliquots before freezing. If aggregation is observed upon thawing, centrifuge at ≥10,000 × g for 10 minutes at 4°C and use only the clarified supernatant. Adding a carrier protein such as 0.1% BSA can also help stabilize the protein at low concentrations during storage.

References

1. Reiter JF, Leroux MR. Genes and molecular pathways underpinning ciliopathies. Nature Reviews Molecular Cell Biology. 2017;18(9):533-547. doi:10.1038/nrm.2017.60
2. Pazour GJ, Agrin N, Leszyk J, Witman GB. Proteomic analysis of a eukaryotic cilium. Journal of Cell Biology. 2005;170(1):103-113. doi:10.1083/jcb.200504008
3. Aladdin Scientific. Recombinant Human CFAP77 Protein, SKU rp374838. Product Datasheet.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.